Solution state NMR structure and dynamics of KpOmpA, a 210 residue transmembrane domain possessing a high potential for immunological applications.
The three-dimensional structure of the outer membrane protein A from Klebsiella pneumoniae transmembrane domain was determined by NMR.This protein induces specific humoral and cytotoxic responses, and is a potent carrier protein. This is one of the largest integral membrane proteins(210 residues) for which nearly complete resonance assignment, including side chains, has been achieved so far. The methodology rested on the use of 900 MHz 3D and 4D TROSY experiments recorded on a uniformly 15N,13C,2H-labeled sample and on a perdeuterated methyl protonated sample. The structure was refined from 920 experimental constraints, giving an ensemble of 20 best structures with an r.m.s. deviation of 0.54 A for the main chain atoms in the core eight-stranded beta-barrel. The protein dynamics was assessed, in a residue-specific manner, by 1H-15N NOEs (pico- to nanosecond timescale), exchange broadening (millisecond to second) and 1H-2H chemical exchange (hour-weeks).
Renault
Marie
M
Université de Toulouse, UPS; Institute of Pharmacology and Structural Biology - IPBS, Toulouse, France
Saurel
Olivier
O
Czaplicki
Jerzy
J
Demange
Pascal
P
Gervais
Virginie
V
Löhr
Frank
F
Réat
Valérie
V
Piotto
Martial
M
Milon
Alain
A
D000595:Amino Acid Sequence; D001425:Bacterial Outer Membrane Proteins / Q000737:chemistry* / Q000276:immunology*; D003902:Detergents / Q000494:pharmacology; D007711:Klebsiella pneumoniae / Q000737:chemistry* / Q000187:drug effects; D009682:Magnetic Resonance Spectroscopy; D008823:Micelles; D008969:Molecular Sequence Data; D010742:Phospholipid Ethers; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D011522:Protons; D012996:Solutions; D013997:Time Factors
C052298:1,2-dihexadecyl-sn-glycero-3-phosphocholine; D001425:Bacterial Outer Membrane Proteins; D003902:Detergents; D008823:Micelles; D010742:Phospholipid Ethers; D011522:Protons; D012996:Solutions; C078167:OMPA outer membrane proteins